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RNA Interference (RNAi) technology has rapidly become one of the key methods used in functional genomics. RNAi is used to block the expression of genes and create phenotypes that can potentially yield clues about the function of these genes. In the postgenomic era, the elucidation of the physiological function of genes has become the rate-limiting step in the quest to develop 'gene-based drugs' and RNAi could potentially play a pivotal role in the validation of such novel drugs. In this 2005 overview, the basic concepts and applications of RNAi biology are discussed. Leading experts from both academia and industry have contributed to this invaluable reference. The volume is foreworded by Andrew Fire, one of the winners of the 2006 Nobel Prize for the discovery of RNA Interference.
Part I. Basic RNAi, siRNA and Gene Silencing Mechanisms
1. RNAi beginnings: overview of the pathway in C. elegans
2. Dicer in RNAi: its roles in vivo and utility in vitro
3. Genes required for RNAi
4. MicroRNAs: a small contribution from worms
5. miRNAs in the brain and the application of RNAi to neurons
Part II. Design and Synthesis of siRNAs
6. Design and synthesis of small interfering RNA (siRNA)
7. Automated design and high throughput synthesis of siRNA
8. Rational design of siRNAs with the Sfold software
9. Enzymatic production of siRNAS
Part III. Vector Development and In Vivo, In Vitro and In Ovo Delivery Methods
10. Six methods of inducing RNAi in mammalian cells
11. Viral delivery of siRNA
12. siRNA delivery by lentiviral vectors: design and applications
13. Liposomal delivery of siRNAs in mice
14. Chemical modifications to achieve increased stability and sensitive detection of siRNA
15. RNAi in postimplantation mouse embryos
16. In ovo RNAi opens new possibilities for functional genomics in vertebrates
Part IV. Gene Silencing in Model Organisms
17. Practical applications of RNAi in C. elegans
18. Inducible RNAi as a forward genetic tool in Trypanosoma brucei
19. RNAi gene silencing in yeast
20. RNA silencing in filamentous fungi: Mucor circinelloides as a model organism
21. RNAi and gene silencing phenomenon mediated by viral suppressors in plants
Part V. Drug Target Validation
22. Delivering siRNA in vivo for functional genomics and novel therapeutics
23. The role of RNAi in drug target validation: application to Hepatitis C
24. RNAi in the drug discovery process
25. RNAi: use in the drug discovery process
Part VI. Therapeutic and Drug Development
26. RNAi-mediated silencing of viral gene expression and replication
27. RNAi in drug development: practical considerations
28. RNAi for the treatment of liver diseases
29. RNAi applications in animal systems
Part VII. Genome Wide RNAi and High Through-Put Analysis
30. High through-put RNAi-by-soaking in Caenorhabditis elegans
31. Tools for integrative genomics: genome-Wide RNAi and expression profiling in Drosophila
32. Microarray analysis and RNA silencing to determine genes functionally important in Mesothelioma
33. High through-put RNAi
34. Generation of highly specific vector-based shRNAi libraries directed against the entire human genome
This book is very timely. The chapters on the basics of RNAi, siRNA and miRNAs are an excellent summary of the recent origins of this topic and the various sections dealing with RNAi technology provided comprehensive background material. I am sure that, even with the rapid progress in RNAi , this book will be useful for both newcomers to the field wishing to get an overview and for experienced researchers looking for a summary of published information. - David Baulcombe The Sainsbury Laboratory, Norwich UK
"Only six years ago Fire and Mello discovered RNAi, as a funny effect in little worms. Now the whole world wants to know about it. In this book Krishnarao Appasani got an all-star crowd of specialists to introduce RNAi: its history, biology, application in genomics research, and possible therapeutic use. Warmly recommended."
- Ronald Plasterk Hubrecht Laboratories, Utrecht, the Netherlands
"A comprehensive treatise on RNA interference has long been needed. That need has now been filled with this book, which places RNAi and its potential squarely in the context of the post human genome era. The successful juxtaposition of the many aspects of RNAi biology and technology should serve to stimulate creative thought across the many diverse disciplines affected by the RNAi revolution."
- Ronald K. Scheule Ph.D., Genzyme Corporation
"This new book is very comprehensive, in many ways picking up where Hannon's RNAi manual leaves off. A good companion to Hannon."
- Frank Slack Ph.D. Yale University
"This extensive collection of timely articles brings us up to date on the current research on RNAi, its science and applications. It is a must read to learn about current status of the art. Its 35 contributions from leading scientists are subdivided into seven sections to facilitate access to the corresponding articles, and cover every aspect of the subject. This is a rapidly advancing field and it is useful for the newcomer both in academia and in industry to obtain a panoramic view and put the current literature in perspective."
- Roberto Weinmann Bristol-Myers Squibb
"This book provides a window to the exciting new RNA interference revolution with special emphasis on applications for biotechnology. It will provide up-to-date background for academic and industry researchers seeking to make use of this powerful research tool."
- Judy Leiberman Harvard Medical School and The CBR Institute for Biomedical Research
'... this most impressive book is not only scientifically informative but also visually appealing. I would have no hesitation recommending this book to anyone interested in RNAi research, particularly newcomers to this daunting area. I am sure this book will become a standard reference point for laboratories working in this area."
"This book provides an excellent, comprehensive description of the current state of the art, in a clear and readily absorbable format, suitable for a wide scientific audience."
- Society for General Microbiology